Removal of Dead Cells from Single Cell Suspensions for Single Cell RNA Sequencing

Demonstrated Protocol, Last Modified on March 31, 2021, Permalink


20200115_CG000093_DeadCell Removal_RevBtoRevC.pdf

A high percentage of non-viable cells may impact the targeted cell recovery in 10x Genomics® Single Cell Protocols. This Demonstrated Protocol outlines best practices for reducing the percentage of non-viable or dead cells from a single cell suspension.

This Protocol was demonstrated using peripheral blood mononuclear cells (PBMCs) and dissociated tissue cells from colorectal cancer (CRC) and clear cell renal carcinoma (CCRC) patients. However, it may be used as a basis for removing dead cells from other cell lines as well as other primary cells in preparation for use in 10x Genomics Single Cell Protocols.

This demonstrated protocol is optimized for counting cells in the range of 700-1200 cells/µl. If using the Single Cell 3' LT v3.1 (low throughput) application, ensure cells are counted as indicated in this protocol and then diluted to the LT specific optimal loading concentration of 100-600 cells/µl using the Cell Dilution Overview in the LT User Guide.